Xiao Lu, Lin Shudian, Xu WenChao, Sun Erwei
Department of Rheumatology and Immunology, The Third Affiliated Hospital, Southern Medical University, Guangzhou, China.
Department of Rheumatology and Immunology, Hainan General Hospital (Hainan Affiliated Hospital of Hainan Medical University), Hainan, China.
Cell Death Discov. 2023 Mar 14;9(1):95. doi: 10.1038/s41420-023-01388-z.
The deposition of monosodium urate (MSU) crystals in arthritic joints of gout seriously damages cartilage. This study aimed to investigate whether MSU crystal-induced cartilage impairment was related to autophagic signaling. mRNAs of cartilage from MSU-induced gouty arthritis rat model were sequenced. MSU crystal-treated human chondrocytes were used to evaluate the function of Sox8. The recombinant Sox8 lentiviral vector (lenti-Sox8) was applied to upregulate the expression of Sox8. Transfection of the mRFP-GFP-LC3 plasmid was evaluated by confocal microscopy. The autophagic vacuoles were stained with monodansylcadaverine and examined by flow cytometry. The morphology of autophagosomes was observed by transmission electron microscopy. The ratio of LC3-II/I in the presence or absence of bafilomycin A1 and the expression levels of Beclin1, Sox8, p-PI3K, PI3K, p-AKT, AKT, p-mTOR, and mTOR were detected by Western blot. In vivo, the effect of Sox8 on cartilage of acute gouty model rats was evaluated by safranin-O/fast green staining and Western blot. The expression of Sox8 was significantly downregulated both in vivo and in vitro. In chondrocytes, MSU crystals reduced the expression of Sox8, inhibited the PI3K/AKT/mTOR signaling pathway, and increased the level of autophagy. Overexpression of Sox8 notably inhibited MSU crystal-induced autophagy by rescuing the phosphorylation levels in the PI3K/AKT/mTOR signaling pathway. In vivo, overexpression of Sox8 remarkably alleviated cartilage damage in acute gouty model rats. These results indicate that downregulation of Sox8 plays an important role in MSU-induced chondrocyte autophagy by modulating PI3K/AKT/mTOR signaling, and overexpression of Sox8 may serve as a novel therapy to prevent the impairment of cartilage in gout arthritis.
尿酸单钠(MSU)晶体在痛风性关节炎关节中的沉积会严重损害软骨。本研究旨在探究MSU晶体诱导的软骨损伤是否与自噬信号传导有关。对MSU诱导的痛风性关节炎大鼠模型的软骨mRNA进行测序。使用MSU晶体处理的人软骨细胞来评估Sox8的功能。应用重组Sox8慢病毒载体(慢病毒-Sox8)上调Sox8的表达。通过共聚焦显微镜评估mRFP-GFP-LC3质粒的转染情况。用单丹磺酰尸胺对自噬泡进行染色,并通过流式细胞术进行检测。通过透射电子显微镜观察自噬体的形态。通过蛋白质免疫印迹法检测在有无巴佛洛霉素A1情况下LC3-II/I的比例以及Beclin1、Sox8、p-PI3K、PI3K、p-AKT、AKT、p-mTOR和mTOR的表达水平。在体内,通过番红O/固绿染色和蛋白质免疫印迹法评估Sox8对急性痛风模型大鼠软骨的影响。Sox8的表达在体内和体外均显著下调。在软骨细胞中,MSU晶体降低了Sox8的表达,抑制了PI3K/AKT/mTOR信号通路,并增加了自噬水平。Sox8的过表达通过挽救PI3K/AKT/mTOR信号通路中的磷酸化水平,显著抑制了MSU晶体诱导的自噬。在体内,Sox8的过表达显著减轻了急性痛风模型大鼠的软骨损伤。这些结果表明,Sox8的下调通过调节PI3K/AKT/mTOR信号传导在MSU诱导的软骨细胞自噬中起重要作用,而Sox8的过表达可能作为一种新的治疗方法来预防痛风性关节炎中的软骨损伤。